CN114438245B - SNP Molecular Markers Linked to the Major QTL Sites for Bacterial Wilt Resistance in Peanut and Its Application - Google Patents
SNP Molecular Markers Linked to the Major QTL Sites for Bacterial Wilt Resistance in Peanut and Its Application Download PDFInfo
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Abstract
本发明涉及分子标记技术领域,具体涉及与花生抗青枯病主效QTL位点连锁的SNP分子标记及其应用。本发明所提供的SNP分子标记BWRB03‑R中SNP多态性位点位于如SEQ ID NO.1所示序列的第201位,多态性为G/A。具体地,SNP分子标记BWRB03‑R中,多态性位点的基因型为A,对应的花生表型为具备较强的青枯病抗性,多态性位点的基因型为G,对应的花生表型为较弱的青枯病抗性。采用本发明提供的SNP分子标记,可以辅助选择抗青枯病的花生材料,有利于推动抗青枯病花生品种的选育,提高育种效率。The invention relates to the technical field of molecular markers, in particular to a SNP molecular marker linked with the main QTL site of peanut resistance to bacterial wilt disease and its application. In the SNP molecular marker BWRB03-R provided by the present invention, the SNP polymorphism site is located at the 201st position of the sequence shown in SEQ ID NO.1, and the polymorphism is G/A. Specifically, in the SNP molecular marker BWRB03-R, the genotype of the polymorphic site is A, the corresponding peanut phenotype has strong bacterial wilt resistance, and the genotype of the polymorphic site is G, corresponding to The peanut phenotype is weaker bacterial wilt resistance. Using the SNP molecular marker provided by the invention can assist in the selection of bacterial wilt-resistant peanut materials, which is beneficial to promote the selection and breeding of bacterial wilt-resistant peanut varieties and improve the breeding efficiency.
Description
技术领域technical field
本发明涉及分子标记技术领域,具体涉及与花生抗青枯病主效 QTL位点qBWRB03连锁的SNP分子标记及其应用。The invention relates to the technical field of molecular markers, in particular to a SNP molecular marker linked with the main QTL site qBWRB03 of peanut resistance to bacterial wilt disease and its application.
背景技术Background technique
花生(Arachis hypogea L.)是重要的油料作物之一。由青枯菌 (Rastoniasolanacearum)引起的青枯病是危害花生生产最重要的细菌性病害。青枯菌从土壤侵入植物根部,沿维管束向上扩展并大量繁殖,最终导致植物导管丧失输水能力而快速枯委死亡。在花生生产中,一般病地的植株病死率为10-30%,重病地的植株病死率可达到80%以上,极端情况下甚至会全部死亡导致绝收。因此,有效地防控青枯病是花生产业发展的重大需求之一。Peanut (Arachis hypogea L.) is one of the important oil crops. The bacterial wilt disease caused by Rastonia solanacearum is the most important bacterial disease affecting peanut production. R. solanacearum invades the roots of plants from the soil, expands upward along the vascular bundles and proliferates, and eventually leads to the loss of water-carrying capacity of plant vessels and rapid withering and death. In peanut production, the mortality rate of plants in general diseased areas is 10-30%, and the mortality rate of plants in severely diseased areas can reach more than 80%. Therefore, effective prevention and control of bacterial wilt disease is one of the major demands for the development of the peanut industry.
选育和利用抗病品种是防治花生青枯病最为经济有效的措施之一。花生对青枯病的抗性表型为数量抗性,发掘和利用稳定的主效 QTL是选育抗青枯病品种的关键基础。通过构建重组自交系群体,在多个环境下对青枯病抗性进行鉴定,结合分子标记基因型进行QTL 分析,可鉴定出稳定表达的主效QTL;利用与QTL连锁的分子标记可以进行分子标记辅助选择,提高育种效率。Breeding and utilizing disease-resistant varieties is one of the most economical and effective measures to control peanut bacterial wilt. The resistance phenotype of peanut to bacterial wilt disease is quantitative resistance, and the discovery and utilization of stable major QTL is the key basis for the selection and breeding of bacterial wilt resistant varieties. By constructing a population of recombinant inbred lines, identifying bacterial wilt resistance in multiple environments, and performing QTL analysis combined with molecular marker genotypes, the major QTLs that are stably expressed can be identified; using molecular markers linked to QTLs can be used to identify major QTLs. Molecular marker-assisted selection to improve breeding efficiency.
目前对花生抗青枯病的QTL定位研究还较少,仅在B02染色体上发掘出了稳定的主效QTL及其分子标记。为了进一步提高青枯病抗性水平,发掘和利用新的抗病QTL主效位点至关重要的。At present, there are few studies on the QTL mapping of peanut resistance to bacterial wilt, and only the stable major QTL and its molecular markers have been discovered on the B02 chromosome. In order to further improve the resistance level of bacterial wilt, it is very important to discover and utilize new QTL major loci for disease resistance.
发明内容SUMMARY OF THE INVENTION
本发明的目的是提供检测花生抗青枯病主效QTL位点qBWRB03 的方法,快速判断花生的青枯病抗性。The purpose of the present invention is to provide a method for detecting the main QTL locus qBWRB03 of peanut resistance to bacterial wilt disease, and to quickly judge the bacterial wilt disease resistance of peanut.
具体地,本发明第一方面,提供一种花生抗青枯病SNP分子标记 BWRB03-R。更具体地,本发明提供的SNP分子标记BWRB03-R与花生抗青枯病主效QTL位点qBWRB03连锁。Specifically, the first aspect of the present invention provides a peanut bacterial wilt resistance SNP molecular marker BWRB03-R. More specifically, the SNP molecular marker BWRB03-R provided by the present invention is linked to the main QTL site qBWRB03 for peanut resistance to bacterial wilt disease.
本发明提供的SNP分子标记BWRB03-R,与位于B03染色体上的抗青枯病主效QTL位点qBWRB03紧密连锁。The SNP molecular marker BWRB03-R provided by the present invention is closely linked to the bacterial wilt resistance major QTL site qBWRB03 located on the B03 chromosome.
本发明提供的SNP分子标记BWRB03-R,SEQ ID NO.1所示序列的第201位存在多态性位点G/A。The SNP molecular marker provided by the present invention is BWRB03-R, and the 201st position of the sequence shown in SEQ ID NO.1 has a polymorphic site G/A.
在所述SNP分子标记BWRB03-R中,具有第201位多态性位点的基因型为A,对应于含有抗青枯病主效QTL位点qBWRB03,基因型为G,对应于不含qBWRB03。In the SNP molecular marker BWRB03-R, the genotype with the 201st polymorphism site is A, which corresponds to the qBWRB03 containing the main QTL for bacterial wilt resistance, and the genotype is G, which corresponds to the absence of qBWRB03 .
本发明提供的SNP分子标记BWRB03-R,使用SEQ ID NO.2-3所示的引物对扩增得到。The SNP molecular marker BWRB03-R provided by the present invention is amplified by using the primer pair shown in SEQ ID NO. 2-3.
第二方面,本发明提供一种引物对,所述引物对的核苷酸序列如 SEQ ID NO.2-3所示。本发明提供的引物对,用于扩增上述SNP分子标记BWRB03-R。In a second aspect, the present invention provides a primer pair, and the nucleotide sequence of the primer pair is shown in SEQ ID NO. 2-3. The primer pair provided by the present invention is used to amplify the above-mentioned SNP molecular marker BWRB03-R.
BWRB03-R的引物对:Primer pair for BWRB03-R:
正向引物序列5’-ATGAAAAGTGAAAAATGAAAAGTGAAAAG GGA-3’(如SEQ ID NO.2所示),Forward primer sequence 5'-ATGAAAAGTGAAAAATGAAAAGTGAAAAG GGA-3' (shown as SEQ ID NO.2),
反向引物序列5’-CAGCGGGGGCAAGTTCAC-3’(如SEQ ID N O.3所示)。Reverse primer sequence 5'-CAGCGGGGGCAAGTTCAC-3' (shown in SEQ ID NO. 3).
第三方面,本发明请求保护含有上述引物对的试剂或试剂盒。In a third aspect, the present invention claims a reagent or kit containing the above-mentioned primer pair.
根据本领域技术人员的理解,本发明还请求保护上述的SNP分子标记BWRB03-R,或如SEQ ID NO.2-3所示的引物对,或含有SEQ ID NO.2-3所述的引物对的试剂或试剂盒,在检测花生抗青枯病主效 QTL位点qBWRB03中的应用。According to the understanding of those skilled in the art, the present invention also claims to protect the above-mentioned SNP molecular marker BWRB03-R, or the primer pair shown in SEQ ID NO.2-3, or the primers containing SEQ ID NO.2-3 The application of the right reagent or kit in the detection of the main QTL locus qBWRB03 in peanut resistance to bacterial wilt.
具体地,第四方面,本发明提供一种检测花生抗青枯病主效QTL 位点qBWRB03的方法,包括:Specifically, in the fourth aspect, the present invention provides a method for detecting the main QTL locus qBWRB03 of peanut resistance to bacterial wilt, comprising:
(1)利用SEQ ID NO.2-3所示引物对,对待测花生的DNA进行 PCR扩增;(1) using the primer pairs shown in SEQ ID NO.2-3 to carry out PCR amplification of the DNA of the peanut to be tested;
(2)分析PCR扩增产物中SNP分子标记BWRB03-R的基因型,根据所述基因型判断待测花生是否含有抗青枯病主效QTL位点 qBWRB03。(2) Analyze the genotype of the SNP molecular marker BWRB03-R in the PCR amplification product, and judge whether the peanut to be tested contains the main QTL site qBWRB03 for resistance to bacterial wilt according to the genotype.
在本发明提供的检测花生抗青枯病主效QTL位点qBWRB03的方法中,若SNP分子标记BWRB03-R的基因型为A,待测花生含有抗青枯病主效QTL位点qBWRB03;若SNP分子标记BWRB03-R的基因型为G,待测花生不含qBWRB03。In the method for detecting the main QTL locus qBWRB03 for resistance to bacterial wilt in peanut provided by the present invention, if the genotype of the SNP molecular marker BWRB03-R is A, the peanut to be tested contains the main QTL locus qBWRB03 for resistance to bacterial wilt; The genotype of the SNP molecular marker BWRB03-R is G, and the peanuts to be tested do not contain qBWRB03.
本发明还请求保护上述的SNP分子标记BWRB03-R,或如SEQ ID NO.2-3所示的引物对,或含有SEQ ID NO.2-3所述的引物对的试剂或试剂盒,在以下任一的应用:The present invention also claims to protect the above-mentioned SNP molecular marker BWRB03-R, or the primer pair shown in SEQ ID NO.2-3, or a reagent or kit containing the primer pair described in SEQ ID NO.2-3, in Application of any of the following:
(1)在花生种质资源鉴定、改良或分子标记辅助育种中的应用;(1) Application in identification, improvement or molecular marker-assisted breeding of peanut germplasm resources;
(2)在花生青枯病抗性早期预测中的应用;(2) Application in early prediction of peanut bacterial wilt resistance;
(3)在筛选抗青枯病花生中的应用。(3) Application in screening peanuts with resistance to bacterial wilt.
第五方面,本发明提供一种鉴定花生青枯病抗性的方法,包括:A fifth aspect, the present invention provides a method for identifying bacterial wilt resistance in peanut, comprising:
(1)利用SEQ ID NO.2-3所示引物对待鉴定花生的DNA进行 PCR扩增;(1) using the primers shown in SEQ ID NO.2-3 to carry out PCR amplification of the DNA of the peanut to be identified;
(2)分析PCR扩增产物中SNP分子标记BWRB03-R的基因型,根据所述基因型判断待鉴定花生的青枯病抗性。(2) Analyze the genotype of the SNP molecular marker BWRB03-R in the PCR amplification product, and determine the bacterial wilt resistance of the peanut to be identified according to the genotype.
在本发明所提供的鉴定花生青枯病抗性的方法中,若SNP分子标记BWRB03-R的基因型为A,待鉴定花生的青枯病抗性强;SNP 分子标记BWRB03-R的基因型为G,待鉴定花生的青枯病抗性弱。In the method for identifying bacterial wilt resistance of peanuts provided by the present invention, if the genotype of the SNP molecular marker BWRB03-R is A, the peanut to be identified has strong bacterial wilt resistance; the genotype of the SNP molecular marker BWRB03-R is strong; For G, the resistance to bacterial wilt of the peanut to be identified is weak.
本发明的有益效果在于:The beneficial effects of the present invention are:
本发明筛选得到与花生抗青枯病主效位点qBWRB03连锁的SNP 分子标记BWRB03-R,并设计得到检测SNP分子标记的引物对,采用花生抗青枯病主效位点qBWRB03的SNP分子标记的检测方法,可以辅助选择抗青枯病的花生材料,有利于推动抗青枯病花生品种的选育,提高育种效率。The present invention obtains the SNP molecular marker BWRB03-R linked with the main locus qBWRB03 of resistance to bacterial wilt disease in peanut, and designs a primer pair for detecting the molecular marker of SNP, using the SNP molecular marker of the main locus qBWRB03 of resistance to bacterial wilt disease in peanut The detection method can assist in the selection of bacterial wilt-resistant peanut materials, which is beneficial to promote the selection and breeding of bacterial wilt-resistant peanut varieties and improve the breeding efficiency.
本发明的SNP分子标记主要是检测是否含有抗青枯病位点 qBWRB03,含有qBWRB03位点的花生材料抵抗青枯病的能力更强。在育种应用中,可以用ICG12625或具有qBWRB03的其他材料做亲本进行杂交,在其后代中会产生含有或不含有qBWRB03的许多个体,通过标记辅助选择保留含有qBWRB03位点的个体材料,再从这些个体材料中再选择产量高、品质优的,最终就可以形成抗青枯病的花生新品种。The SNP molecular marker of the present invention mainly detects whether it contains the bacterial wilt disease resistance site qBWRB03, and the peanut material containing the qBWRB03 site has stronger resistance to bacterial wilt disease. In breeding applications, ICG12625 or other materials with qBWRB03 can be used as parents for crosses, and many individuals with or without qBWRB03 will be produced in their progeny, and individual materials containing the qBWRB03 locus are retained through marker-assisted selection, and then from these If the individual materials are selected with high yield and good quality, new peanut varieties with resistance to bacterial wilt can finally be formed.
附图说明Description of drawings
图1为为本发明中花生抗青枯病主效位点qBWRB03在染色体B03 的定位图,其中纵向垂直线表示花生染色体B03,B03右侧横向短线段表示染色体上的遗传重组bin;左侧横向短线段表示bin间的遗传距离(cM);抗青枯病主效位点qBWRB03定位于bin位点c13b013和c13b014之间1cM的区域,B03右侧列出了该QTL在2019~2020两年间检测到的置信区间。Fig. 1 is the localization map of peanut bacterial wilt resistance major locus qBWRB03 on chromosome B03 in the present invention, wherein the vertical vertical line represents peanut chromosome B03, and the horizontal short line segment on the right side of B03 represents the genetic recombination bin on the chromosome; The short line segment represents the genetic distance (cM) between bins; the main locus of bacterial wilt resistance qBWRB03 is located in the 1cM region between the bin loci c13b013 and c13b014, and the right side of B03 lists the QTL detected in 2019-2020 to the confidence interval.
具体实施方式Detailed ways
下面将结合实施例对本发明的优选实施方式进行详细说明。需要理解的是以下实施例的给出仅是为了起到说明的目的,并不是用于对本发明的范围进行限制。本领域的技术人员在不背离本发明的宗旨和精神的情况下,可以对本发明进行各种修改和替换。The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples are given for illustrative purposes only, and are not intended to limit the scope of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from the spirit and spirit of the present invention.
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.
实施例1Example 1
本实施例提供与花生抗青枯病主效QTL位点qBWRB03连锁的 SNP分子标记BWRB03-R的开发过程,如下。This example provides the development process of the SNP molecular marker BWRB03-R linked to the major QTL site qBWRB03 for resistance to bacterial wilt in peanut, as follows.
供试材料:以花生感青枯病品种中花10为母本,花生抗青枯病种质ICG12625为父本进行杂交,通过单粒传法获得包含140个株系的重组自交系(RIL)群体。Materials to be tested: crossed with the peanut germplasm susceptible to bacterial wilt disease Zhonghua 10 as the female parent and the peanut germplasm resistant to bacterial wilt disease ICG12625 as the male parent, and obtained a recombinant inbred line (RIL) containing 140 lines by single seed propagation. )group.
表现型鉴定:2019年和2020年在中国农业科学院油料作物研究所红安青枯病病圃鉴定了中花10、ICG12625及RIL群体140个株系的青枯病抗性。采用完全随机区组实验设计,3次重复。每次重复每份材料种植1行20株,行距30厘米,株距10厘米。采取标准的田间管理方式。全部出苗后,统计每份材料的总株数。在青枯病开始发病后,统计病死株数,直至收获。然后计算成活率:(总株数-病死株数)/总株数×100%,成活率越高的材料对青枯病的抗性越好。Phenotypic identification: In 2019 and 2020, the bacterial wilt resistance of 140 lines of Zhonghua 10, ICG12625 and RIL populations were identified in the Hong'an bacterial wilt disease nursery of the Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences. A completely randomized block experimental design was used with 3 replicates. Each repetition of each material was planted with 20 plants in a row, with a row spacing of 30 cm and a plant spacing of 10 cm. Standard field management methods were adopted. After all seedlings emerged, the total number of plants in each material was counted. After the onset of bacterial wilt disease, the number of dead plants was counted until harvest. Then calculate the survival rate: (total number of plants - number of dead plants)/total number of plants × 100%, the higher the survival rate of materials, the better the resistance to bacterial wilt.
遗传连锁图谱的构建:用CTAB法提取亲本和RIL群体140个单株叶片DNA;每个样本DNA建一个300-500bp Illumina Paired-end(PE) 文库,进行PE150测序。下机的原始数据进行质控后得到高质量的cl eandata,然后使用BWA软件将cleandata比对到四倍体栽培花生参考基因组上(https://www.peanutbase.org/data/public/Arachis_hypogaea/Tifrunner.gnm1.KYV3/),最后使用GATK的HaplotypeCaller模块进行SNP 检测。将基因型相同的SNP位点合并成bins,最后计算bins遗传图距从而得到终连锁图谱。该连锁图谱包含2701个bin位点,总长度为1469. 56cM,标记间平均距离为0.54cM。Construction of genetic linkage map: DNA from 140 individual leaves of parental and RIL populations was extracted by CTAB method; a 300-500bp Illumina Paired-end (PE) library was constructed for each sample DNA, and PE150 sequencing was performed. The raw data from the machine is quality controlled to obtain high-quality cleandata, and then BWA software is used to compare the cleandata to the reference genome of tetraploid cultivated peanut (https://www.peanutbase.org/data/public/Arachis_hypogaea/ Tifrunner.gnm1.KYV3/), and finally use the HaplotypeCaller module of GATK for SNP detection. The SNP sites with the same genotype were merged into bins, and finally the genetic map distance of bins was calculated to obtain the final linkage map. The linkage map contains 2701 bin sites, the total length is 1469.56cM, and the average distance between markers is 0.54cM.
QTL的定位:利用WinQTLCart软件的区间作图法开展成活率的Q TL分析,两个环境的成活率表型数据均在染色体B03上c13b013~c13b 014区间检测到主效QTL位点qBWRB03,主效QTL位点qBWRB03在两个环境的LOD值分别为4.82和5.11,表型解释率分别为12.01%和10.5 8%。Mapping of QTL: The QTL analysis of the survival rate was carried out using the interval mapping method of WinQTLCart software. The survival rate phenotype data of the two environments were all detected in the c13b013~c13b014 interval on chromosome B03. The main QTL locus qBWRB03, the main effect The LOD values of the QTL locus qBWRB03 in the two environments were 4.82 and 5.11, respectively, and the phenotypic interpretation rates were 12.01% and 10.58%, respectively.
两个环境的置信区间相互重叠,位于一个1cM的区间(6.7~7.7 cM),其位置的示意图见图1。因此,花生抗青枯病主效位点qBWRB 03能在两年实验中稳定表达,其抗性等位基因来源于ICG12625。The confidence intervals for the two environments overlapped each other and were located in a 1 cM interval (6.7–7.7 cM), a schematic diagram of which is shown in Figure 1. Therefore, the major locus of bacterial wilt resistance in peanut, qBWRB 03, can be stably expressed in the two-year experiment, and its resistance allele is derived from ICG12625.
利用c13b013~c13b014区间中的一个SNP(位于上述参考基因组B 03染色体125,198,196bp(G/A变异))开发出一个SNP分子标记BWR B03-R(SEQ ID NO.1),扩增产物二代高通量测序检测SNP位点碱基组成:在ICG12625中检测出A碱基,属于含有qBWRB03抗青枯病等位基因;而在中花10号中检测出G碱基,则不含有qBWRB03抗青枯病等位基因。A SNP molecular marker BWR B03-R (SEQ ID NO. 1) was developed by using a SNP in the interval c13b013~c13b014 (located on chromosome 125, 198, 196 bp (G/A variation) of the above reference genome B 03), and the amplified product was second-generation high The base composition of SNP sites was detected by flux sequencing: the A base was detected in ICG12625, which belonged to the allele containing qBWRB03 resistance to bacterial wilt; while the G base was detected in Zhonghua No. 10, it did not contain qBWRB03 anti-green wilt allele Blight allele.
针对实施例1提供的分子标记BWRB03-R,设计引物如SEQ ID NO.2-3所示。For the molecular marker BWRB03-R provided in Example 1, the designed primers are shown in SEQ ID NO. 2-3.
实施例2Example 2
本实施例提供,利用与抗青枯病主效位点qBWRB03相连锁的SNP 分子标记BWRB03-R,对实施例1中两个环境均获得成活率数据的 ICG12625和中花10号的131个RIL进行检测。具体步骤如下。This example provides, using the SNP molecular marker BWRB03-R linked to the main locus of bacterial wilt resistance qBWRB03, for ICG12625 and 131 RILs of Zhonghua No. 10 for which survival rate data were obtained in both environments in Example 1 test. Specific steps are as follows.
以基因组DNA为模板,SEQ ID NO.2-3所示序列为引物,用 KAPA2G FastMultiplex Mix试剂盒扩增SNP分子标记BWRB03-R。 PCR条件为:95℃预变性3min;95℃变性15s、55℃复性30s、72℃延伸30s,共30个循环;最后72℃延伸5min,4℃保温。扩增产物添加测序接头后用Illumina HiSeq平台进行Paired-end 150bp(PE150) 测序,测序序列比对到参考序列,检测SNP位点的碱基。如果RIL 与ICG12625的扩增片段检测出的SNP位点碱基一致,说明该株系含有qBWRB03的抗青枯病等位基因。同时用实际测得的成活率(测试方法见实施例1)结果与分子标记检测结果进行验证。Using genomic DNA as a template and the sequences shown in SEQ ID NO. 2-3 as primers, the SNP molecular marker BWRB03-R was amplified with the KAPA2G FastMultiplex Mix kit. PCR conditions were: pre-denaturation at 95 °C for 3 min; denaturation at 95 °C for 15 s, renaturation at 55 °C for 30 s, extension at 72 °C for 30 s, a total of 30 cycles; final extension at 72 °C for 5 min, and incubation at 4 °C. After adding sequencing adapters to the amplified products, Paired-end 150bp (PE150) sequencing was performed on the Illumina HiSeq platform, and the sequenced sequences were compared to the reference sequences, and the bases of the SNP sites were detected. If the bases of RIL and the SNP site detected by the amplified fragment of ICG12625 are consistent, it indicates that the strain contains the bacterial wilt resistance allele of qBWRB03. At the same time, the results of the actually measured survival rate (see Example 1 for the test method) and the detection results of molecular markers were used for verification.
结果显示,在131个RIL中,62个RIL的SNP分子标记基因型与ICG12625相同(基因型分类1,A),说明含有纯合的主效抗青枯病QTL位点qBWRB03;而69个RIL的SNP分子标记基因型与中花10号相同(基因型分类2,G),说明不含有qBWRB03。The results showed that among the 131 RILs, the SNP molecular marker genotype of 62 RILs was the same as that of ICG12625 (genotype classification 1, A), indicating that they contained a homozygous major anti-bacterial wilt QTL locus qBWRB03; The genotype of the SNP molecular marker is the same as that of Zhonghua No. 10 (genotype classification 2, G), indicating that it does not contain qBWRB03.
T检验发现,含有qBWRB03的62个RIL在2019和2020两年鉴定的成活率中均显著高于不含有qBWRB03的69个RIL(表1)。因此,同时用本发明的与抗青枯病主效位点qBWRB03相连锁的SNP 分子标记BWRB03-R预测花生对青枯病抗性有较好的预测效果。The T-test found that the 62 RILs containing qBWRB03 had significantly higher survival rates identified in both 2019 and 2020 than the 69 RILs that did not contain qBWRB03 (Table 1). Therefore, using the SNP molecular marker BWRB03-R linked with the main locus of bacterial wilt resistance qBWRB03 of the present invention to predict the resistance of peanut to bacterial wilt disease has a good prediction effect.
表1利用本发明的SNP分子标记进行选择RIL株系的成活率差异Table 1 Use the SNP molecular marker of the present invention to select the difference in survival rate of RIL strains
**表示在0.01水平上,基因型分类1与基因型分类2之间存在显著差异。基因型分类1的基因型与ICG12625相同,基因型分类2的基因型与中花10号相同。** Indicates a significant difference between genotype class 1 and genotype class 2 at the 0.01 level. The genotype of genotype classification 1 is the same as that of ICG12625, and the genotype of genotype classification 2 is the same as that of Zhonghua 10.
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。Although the present invention has been described in detail above with general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention fall within the scope of the claimed protection of the present invention.
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